Attenuated <i>T</i> <sub>2</sub> relaxation by mutual cancellation of dipole–dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solution

Proceedings of the National Academy of Sciences of the United States of America - Tập 94 Số 23 - Trang 12366-12371 - 1997
Konstantin Pervushin1, Roland Riek1, Gerhard Wider1, Kurt Wüthrich1
1Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Hönggerberg, CH-8093 Zurich, Switzerland

Tóm tắt

Fast transverse relaxation of 1 H, 15 N, and 13 C by dipole-dipole coupling (DD) and chemical shift anisotropy (CSA) modulated by rotational molecular motions has a dominant impact on the size limit for biomacromolecular structures that can be studied by NMR spectroscopy in solution. Transverse relaxation-optimized spectroscopy (TROSY) is an approach for suppression of transverse relaxation in multidimensional NMR experiments, which is based on constructive use of interference between DD coupling and CSA. For example, a TROSY-type two-dimensional 1 H, 15 N-correlation experiment with a uniformly 15 N-labeled protein in a DNA complex of molecular mass 17 kDa at a 1 H frequency of 750 MHz showed that 15 N relaxation during 15 N chemical shift evolution and 1 H N relaxation during signal acquisition both are significantly reduced by mutual compensation of the DD and CSA interactions. The reduction of the linewidths when compared with a conventional two-dimensional 1 H, 15 N-correlation experiment was 60% and 40%, respectively, and the residual linewidths were 5 Hz for 15 N and 15 Hz for 1 H N at 4°C. Because the ratio of the DD and CSA relaxation rates is nearly independent of the molecular size, a similar percentagewise reduction of the overall transverse relaxation rates is expected for larger proteins. For a 15 N-labeled protein of 150 kDa at 750 MHz and 20°C one predicts residual linewidths of 10 Hz for 15 N and 45 Hz for 1 H N , and for the corresponding uniformly 15 N, 2 H-labeled protein the residual linewidths are predicted to be smaller than 5 Hz and 15 Hz, respectively. The TROSY principle should benefit a variety of multidimensional solution NMR experiments, especially with future use of yet somewhat higher polarizing magnetic fields than are presently available, and thus largely eliminate one of the key factors that limit work with larger molecules.

Từ khóa


Tài liệu tham khảo

10.1126/science.165.3890.251

10.1016/S0076-6879(94)39017-7

10.1051/epn/19861701011

10.1142/2732

10.1107/S0907444994010188

10.1021/ja960627a

10.1007/BF00176005

10.1021/ja952207b

10.1021/ja00363a009

10.1021/ja960510m

T C Farrar, T C Stringfellow Encyclopedia of NMR, eds D M Grant, R K Harris (Wiley, New York) 6, 4101–4107 (1996).

10.1016/0079-6565(78)80004-1

R R Ernst, G Bodenhausen, A Wokaun The Principles of Nuclear Magnetic Resonance in One and Two Dimensions (Clarendon, Oxford, 1987).

A Abragam The Principles of Nuclear Magnetism (Clarendon, Oxford, 1961).

M Goldman J Magn Reson 60, 437–452 (1984).

R E London J Magn Reson 86, 410–415 (1990).

J W Peng, G Wagner J Magn Reson 98, 308–332 (1992).

10.1002/j.1460-2075.1990.tb07617.x

10.1006/jmbi.1994.1296

10.1021/bi00449a003

10.1021/bi00185a040

10.1021/ja00510a007

10.1016/0009-2614(80)80041-8

10.1016/0079-6565(76)80001-5

10.1080/00268979200100511

L E Kay, L K Nicholson, F Delaglio, A Bax, D A Torchia J Magn Reson 97, 359–375 (1992).

10.1006/jmrb.1996.0057

10.1021/ja00216a006

10.1017/S0033583500004029

10.1021/ja00124a014

10.1016/0079-6565(84)80005-9

10.1006/jmra.1993.1016

10.1006/jmre.1997.1161

10.1007/BF02192855

10.1063/1.1743033

10.1063/1.1725007

10.1021/ja970876e

Thông tin
Thông tin xuất bản

Proceedings of the National Academy of Sciences of the United States of America

Tập 94 Số 23

12366-12371

Thông tin tác giả